Isolation, characterization and localization of A 45 000 molecular weight, soluble glycoprotein from the lung in pulmonary alveolar proteinosis

Robert H. Glew , John U. Balis , Sue Shelley , Theresa Kuhlenschmidt , Ronald Jaffe
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引用次数: 5

Abstract

A carbohydrate-rich, water-soluble glycoprotein has been isolated in pure form from delipidated lung lavage fluid from a patient with pulmonary alveolar proteinosis, in a three-step procedure involving ion-exchange and gel filtration chromatography. The molecular weight of the glycoprotein was determined to be 45 900 by sedimentation equilibrium analysis in the analytical ultracentrifuge and 45 000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, indicating a single polypeptide chain. Nearly half of the mass of the glycoprotein is comprised of carbohydrate that is contributed by 24 residues sialic acid, 23 residues N-acetylglucosamine, 6 residues N-acetylgalactosamine, 19 residues galactose, 4 residues mannose, 1 residue fucose and 1 residue glucose per mol. Unlike a number of collagen-related glycoproteins that have been isolated by others from insoluble lung contents in pulmonary proteinosis, the water-soluble glycoprotein described in the present report does not contain hydroxyproline or hydroxylysine and contains less than 10% of its amino acid residues as glycine. Using rabbit antibodies directed against our purest preparation of material and an immunoperoxidase staining procedure, the 45 000 molecular weight glycoprotein was localized to the thin film of fluid lining the surfaces of alveoli in normal human lungs.

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肺泡蛋白沉积症患者肺可溶性糖蛋白A的分离、鉴定和定位
从肺泡蛋白沉积症患者的脱水肺灌洗液中分离出一种富含碳水化合物的水溶性糖蛋白,采用离子交换和凝胶过滤色谱法进行三步分离。在十二烷基硫酸钠存在下,经聚丙烯酰胺凝胶电泳测定糖蛋白分子量为45 000,经分析超离心机沉淀平衡分析测定糖蛋白分子量为45 900,为单链多肽。糖蛋白近一半的质量是由碳水化合物组成的,每摩尔有24个唾液酸残基、23个n-乙酰氨基葡萄糖残基、6个n-乙酰半乳糖残基、19个半乳糖残基、4个甘露糖残基、1个焦糖残基和1个葡萄糖残基。与其他一些从肺蛋白沉积症的不溶性肺内容物中分离出来的胶原蛋白相关糖蛋白不同,本报告所述的水溶性糖蛋白不含羟脯氨酸或羟赖氨酸,其氨基酸残基中甘氨酸的含量少于10%。利用兔抗体和免疫过氧化物酶染色方法,将45,000分子量的糖蛋白定位在正常人肺肺泡表面的液体薄膜上。
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